Zhen Li, Lina Sun, Yulu Wang, Bolin Liu and Fengjiao Xin*,
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引用次数: 0
摘要
香兰素是世界上最广泛使用的调味剂之一,具有很高的应用价值。然而,由于底物吸收效率低以及香兰素对细胞生长的抑制作用,香兰素生物合成的产量仍然有限。在此,我们通过在生产香兰素的工程大肠杆菌菌株 VA 中过表达编码候选转运体的基因,筛选出了高效阿魏酸导入体 TodX 和香兰素导出体 PP_0178 和 PP_0179,并通过在香兰素自诱导启动子 ADH7 上共表达 TodX 和 PP_0178/PP_0179 进一步构建了自调节双向转运系统。与 VA 菌株相比,VA-TodX-PP_0179 菌株能高效地将阿魏酸跨细胞膜转运并转化为香兰素,从而显著提高了底物利用率(14.86%)和香兰素滴度(51.07%)。这项研究表明,自律性双向转运系统能显著提高底物吸收效率,同时缓解香兰素毒性问题,为香兰素的生物合成提供了一条可行的途径。
Construction of a Novel Vanillin-Induced Autoregulating Bidirectional Transport System in a Vanillin-Producing E. coli Cell Factory
Vanillin is one of the world’s most extensively used flavoring agents with high application value. However, the yield of vanillin biosynthesis remains limited due to the low efficiency of substrate uptake and the inhibitory effect on cell growth caused by vanillin. Here, we screened high-efficiency ferulic acid importer TodX and vanillin exporters PP_0178 and PP_0179 by overexpressing genes encoding candidate transporters in a vanillin-producing engineered Escherichia coli strain VA and further constructed an autoregulatory bidirectional transport system by coexpressing TodX and PP_0178/PP_0179 with a vanillin self-inducible promoter ADH7. Compared with strain VA, strain VA-TodX-PP_0179 can efficiently transport ferulic acid across the cell membrane and convert it to vanillin, which significantly increases the substrate utilization rate efficiency (14.86%) and vanillin titer (51.07%). This study demonstrated that the autoregulatory bidirectional transport system significantly enhances the substrate uptake efficiency while alleviating the vanillin toxicity issue, providing a promising viable route for vanillin biosynthesis.
期刊介绍:
The Journal of Agricultural and Food Chemistry publishes high-quality, cutting edge original research representing complete studies and research advances dealing with the chemistry and biochemistry of agriculture and food. The Journal also encourages papers with chemistry and/or biochemistry as a major component combined with biological/sensory/nutritional/toxicological evaluation related to agriculture and/or food.